Lin Hong1,2, Stephanie Chavez1, Yelena Smagley1, Alexandre Chigaev1, Larry A Sklar1,2,3. 1. Department of Pathology, The University of New Mexico, Albuquerque, New Mexico, 87131. 2. Department of Pathology, Center for Molecular Discovery, The University of New Mexico, Albuquerque, New Mexico, 87131. 3. Cancer Research and Treatment Center, The University of New Mexico, Albuquerque, New Mexico, 87131.
Abstract
BACKGROUND: Cdc42 GTPase has important roles in regulating intracellular actin reorganization. The current methods to monitor actin changes are typically complex and point by point. METHODS: The effects of Cdc42 inhibitors on the side scatter changes were tested in a newly developed continuous assay using the flow cytometer. Staining with fluorescently labeled phalloidin was used for comparison. RESULTS: Cdc42-specific inhibitors caused dose-dependent changes of both the right-angle side scatter and the phalloidin-stained actin. CONCLUSIONS: The right-angle light scatter change can be used as a method to circumvent phalloidin staining and be an early convenient step in screening Cdc42 inhibitors.
BACKGROUND:Cdc42 GTPase has important roles in regulating intracellular actin reorganization. The current methods to monitor actin changes are typically complex and point by point. METHODS: The effects of Cdc42 inhibitors on the side scatter changes were tested in a newly developed continuous assay using the flow cytometer. Staining with fluorescently labeled phalloidin was used for comparison. RESULTS:Cdc42-specific inhibitors caused dose-dependent changes of both the right-angle side scatter and the phalloidin-stained actin. CONCLUSIONS: The right-angle light scatter change can be used as a method to circumvent phalloidin staining and be an early convenient step in screening Cdc42 inhibitors.